Process for preparation of a solid phase system

ABSTRACT

The invention provides a method for the production of a solid phase system of the formula F--(--R 3  --Y--CO--CR 2  ═CR 1  --CH 2  --X) m  using new allyl esters of the formula: 
     
         X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --A  (I). 
    
     R 1  and R 2  are hydrogen, halogen, alkyl or aryl, R 3  is a spacer, X is acyloxy where acyl is the residue of an aliphatic carboxylic acid or is RCO-- in which R is an organic group. Y is oxygen or sulphur or an --NH-- group and A is a grouping which can react with a solid carrier material F, which contains functional groups B. A and B are groups which react with one another with condensation and/or addition and formation of a linkage between the solid carrier material and X--CH 2  --CR 1  ═CR 2  --CO--Y--R 3  --.

This is a divisional application of application Ser. No. 07/576,366,filed Aug. 31, 1990, and now U.S. Pat. No. 5,214,195.

The present invention is concerned with new allyl esters, processes forthe preparation thereof and the use thereof for the synthesis of solidphase systems for solid phase reactions and especially for the solidphase synthesis of peptides, glycopeptides and proteins.

Having regard to the increasing requirements for pharmaceuticals,foodstuff additives and other active materials with regard to theselectivity of the action, the compatibility and the biologicaldegradability, the precision synthesis of peptides is again of greatimportance. In spite of the now highly developed genetechnologicaltechniques, this also applies to the chemical synthesis of peptides bywhich alone, for example, peptides with non-natural constructional unitsand structural elements are accessible.

For the chemical synthesis of peptides, the introduction of the solidphase synthesis according to R. B. Merrifield (see R. B. Merrifield,J.A.C.S., 85, 2149/1963) signifies a great advance. This applies inspite of the problems which have, in the meantime, been recognized whichoccur ever again in the case of these solid phase peptide syntheses withregard to the purity of the synthesised products.

In the solid phase syntheses, as C-terminal protective and anchor groupsthere serve benzyl esters which permit the splitting off of the built-uppeptides from the polymeric carrier under more or less strongly acidicconditions. The acidic splitting off conditions--in the classicalMerrifield synthesis use was made of hydrogen bromide in varioussolvents or hydrogen fluoride--have the disadvantage that undesired sidereactions, such transpeptidisations or transalkylations, can therebyoccur. The synthesis of glycopeptides can scarcely be carried out inthis way because the sensitive glycosidic bonds of these molecules arecleaved or anomerised under acidic conditions (with regard to the priorart of glycopeptide synthesis, cf. for example, H. Kunz, AngewandteChemie, 99/1987; Angewandte Chemie, Int. Engl. Ed., 26, 294/1987).

It is an object of the present invention to provide solid phase systems,especially for the solid phase synthesis of peptides and glycopeptides,in the case of which the peptides and glycopeptides can be split offfrom the polymeric carrier selectively and under such conditions that nocleavage of the glycosidic bonds and no anomerisations take place.

From published Federal Republic of Germany Patent Specification No. 3803 545, there is known an allylic side chain-containing solid phasesystem in which the allylic side chains are bound to a solid carriermaterial. The C-terminal protective (anchor) groups used in these solidphase systems, via which the amino acids and peptides, glycopeptides andthe like can be bound to the polymeric carrier, belong to the allylester type. Allyl esters can, as C-terminal protective groups inglycopeptide, nucleotide and peptide syntheses, be split off selectivelyand under mild, almost neutral conditions from the blocked compounds(cf. H. Kunz, Angew. Chemie, 99, 297/1987). This is achieved by noblemetal catalysis, for example by catalysis with compounds of the platinummetal group, such as ruthenium, rhodium, palladium, osmium, iridium andplatinum, and especially by catalysis with rhodium (I) compounds (cf. H.Waldmann, H. Kunz, Liebigs Ann. Chem., 1983, 1712) or with reactionscatalysed by palladium (O) compounds (H. Kunz, H. Waldmann, Angew.Chemie, 96, 47/1984; Angew. Chemie Int. Engl. Ed., 23, 71/1984; H. Kunzand C. Unverzagt, Angew. Chemie, 96, 426/1984; Angew. Chemie, Int. Engl.Ed., 23, 436/1984). By the transfer of the deblocking method to thesolid phase peptide synthesis, it is achieved that the removal of thepeptide, glycopeptide or nucleotide chain built up on the carrier ispossible practically under neutral conditions.

We have now found that allylic side chain-containing solid phasesystems, such as are described in published Federal Republic of GermanyPatent Specification No. 38 03 545, can also be built up when aconjugate with an N-protected amino acid is first prepared in solutionwhich, on its carboxyl function, carries the allylic principle and thisconjugate is then elongated with a spacer grouping which terminates witha functional group for anchoring on to a solid carrier. These "tripleconstructions" (as it were "preformed terminal handles"), which can besynthesised conventionally and in a simple way in solution, which areanalytically exactly definable compounds, can now be connected with thesolid carrier with the formation of a solid phase system of the typedescribed in published Federal Republic of Germany Patent SpecificationNo. 38 03 545.

Thus, according to the present invention, there are provided new allylesters of the general formula:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --A  (I)

wherein R¹ and R², which can be the same or different, are hydrogen orhalogen atoms or alkyl or aryl radicals, R³ is a linking grouping(spacer), X is an acyloxy group, whereby acyl in the acyloxy radical isthe residue of a carboxylic acid or a radical RCO--, in which R is anorganic radical, Y is an oxygen or sulphur atom or, especially, an--NH-- group and A is a grouping which can react with a solid carriermaterial F, which contains appropriate functional groups B, either afterselective deblocking or directly with the atom grouping B withcondensation and/or addition and formation of a linkage between thesolid carrier material and the radical X--CH₂ --CR¹ ═CR² --CO--Y--R³ --.

Preferred compounds of general formula (I) are those in which the acylradical RCO-- is a protected or unprotected residue of an amino acid,peptide, glycopeptide, nucleotide, hydroxycarboxylic acid, dicarboxylicacid or tricarboxylic acid, the acyl radical more preferably being theresidue of an N-protected amino acid.

Preferred allyl esters of general formula (I) are also those in which R¹and R² are hydrogen atoms.

Also preferred are compounds of general formula I, wherein the grouping--R³ --A, besides the intact grouping X, is a selectively deblockablecarboxylic acid ester grouping, for example --CH₂ --CH₂ --COO--CH₂ --CH₂--Br, of the radical --CH₂ --CH₂ --COOH.

The alkyl radicals R¹ and R² can be straight-chained or branched, thealkyl radicals preferably containing up to 7 and more preferably up to 3carbon atoms.

An aryl radical R¹ or R² is preferably an unsubstituted or substitutedmono- or dicyclic aryl radical, for example a naphthyl-1 or -2 radicalor especially a phenyl radical. The aryl radical can contain one or moresubstituents, for example lower alkyl radicals and/or halogen atoms, butis preferably unsubstituted. Two alkyl substituents can, together withthe aryl radical, also form a system of two or more rings, for exampletetrahydronaphthalene.

The radical R³ is a linking group (spacer or linker) and can be, forexample, one of the spacer groupings previously used in the solid phasetechnique (Merrifield technique) (for example CASET or CAMET; cf. RomppsChemie-Lexikon, 8th edition, page 2543). The nature of the group R³thereby depends especially also upon the process, described hereinafterin more detail, for the preparation of the solid phase systems of thegeneral formula (II) and especially upon the nature of the carriermaterial and upon the functional groups A and B present on the carriermaterial and in the compounds of general formula (I). The group R³ canbe, for example, an alkylene, aralkylene or arylene radical in which oneor more --CH₂ -- groups can be substituted and/or replaced byheteroatoms. R³ is preferably the grouping --(CH₂)_(n) --COO--CH₂ --CH₂-- or --(CH₂)_(n) --, wherein n is at least 2, more preferably 2 to 10and most preferably 2 to 5.

In an acyloxy radical X, in which acyl is the residue of an aliphaticcarboxylic acid, acyl is preferably the acid residue of an aliphaticcarboxylic acid containing up to 7 and preferably up to 4 carbon atoms,for example a formyl, acetyl or propionyl radical.

An acyl radical RCO-- is preferably the unprotected and more preferablythe protected residue of an amino acid, of a peptide, glycopeptide,nucleotide, hydroxycarboxylic acid, dicarboxylic acid or tricarboxylicacid and, in particular, is the residue of an N-protected amino acid.

In the compounds of general formula (I), A is preferably a carboxylicacid ester group which, in the presence of the intact amino protectivegroup, can be split into X or RCO--, for example a methyl ester,tert.-butyl ester, 2-haloethyl ester, allyl ester, fluorenyl-9-methylester or an active ester or is itself of carboxyl group. Thus, ingeneral formula (I), the R³ --A-- grouping is preferably, for example,--CH₂ --CH₂ --COO--CH₂ --CH₂ --Br or --CH₂ --CH₂ --COOH.

For the solid carrier material, it is preferable to start from one whichcontains functional groups B which are well suited for the reaction withthe residues A of the compounds of general formula (I).

The carrier material is preferably an organic or inorganic polymer, forexample a synthetic, semisynthetic or natural polymer. Such polymersinclude, for example, cross-linked polyacrylamides, methacrylates,dextrans (for example Sephadex^(R)), cellulose and, in particular,polystyrene. However, the carrier material can also consist of a solidbase material which is coated with a material appropriate for thelinkage with the allyl side chains, for example is coated with anappropriate polymer or a cross-linked protein. The base material can be,for example, glass, silica gel or also a synthetic resin. Organicpolymers for the carrier material and suitable as coatings include, forexample, polyacrylamides, polyethylene glycols and especiallypolystyrenes. As solid, functional group B-containing carrier material,there is particularly used an aminomethylated polystyrene.

The residues A and B are those groupings which react either directly orafter selective deblocking with condensation and/or addition andformation of a linkage between the carrier material and the radicalX--CH₂ --CR¹ ═CR² --CO--Y--R³ --. Such groups are preferably thoseusually employed in condensation and addition reactions, for exampleamino groups such as in the form of an aminomethyl radical on the solidcarrier, halogen atoms, ester groupings, carboxyl groups, nitrile groupsand the like. As a rule, the condensation and/or addition reactions canbe carried out in known manner, for example with the splitting off ofwater, halogen, hydrogen or the like. The residue A is preferably ahalogen atom or a carboxyl group.

Halogen is preferably fluorine, chlorine, bromine or iodine. It ispreferably chlorine or bromine and, in the meaning of A, is especiallybromine.

In a preferred embodiment for the build up the solid phase system, forexample an aminomethyl radical (group B)-containing polystyrene isreacted with a compound of general formula (I) and especially with acompound of general formula (I) in which R³ --A-- is the grouping --CH₂--CH₂ --COOH which is produced from an appropriate selectively-cleavableester, for example --CH₂ --CH₂ --COO--CH₂ --CH₂ Br.

The present invention also provides a process for the preparation of thecompounds of general formula (I) according to the present invention,wherein

a) a compound of the general formula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --COOCH.sub.2 CCl.sub.3( 2)

in which R¹ and R² have the above-given meanings, is reacted with a saltof an acid of the general formula XH (1), in which X has the above-givenmeaning, the so-obtained trichloroethyl ester of the general formula:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --COOCH.sub.2 CCl.sub.3( 3)

in which R¹, R² and X have the above-given meanings, is converted intothe free acid of the general formula:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --COOH                (4)

in which R¹, R² and X have the above-given meanings, and this is reactedwith a compound of the general formula:

    HY--R.sup.3 --A                                            (5)

in which A, R³ and Y have the above-given meanings, to give a compoundof general formula (I); or

b) a compound of the general formula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --COOH                 (6)

in which R¹ and R² have the above-given meanings, is reacted with acompound of general formula (5) to give a compound of the generalformula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --A   (7)

in which A, R¹, R² and Y have the above-given meanings, and thiscompound is then converted into a compound of general formula (I) byreaction with a salt of an acid of general formula XH (1), in which Xhas the above-given meaning; or

c) a compound of the general formula (6) is reacted with a compound ofthe general formula:

    HY--R.sup.3 --COO--CH.sub.2 --CH═CH.sub.2              ( 8)

in which R³ and Y have the above-given meanings, to give a compound ofthe general formula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --COO--CH.sub.2 --CH═CH.sub.2                                         ( 9)

in which R¹, R², R³ and Y have the above-given meanings, this compoundis then reacted with the salt of an acid of the general formula XH (1),in which X has the above-given meaning, to give a compound of thegeneral formula:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --COO--CH.sub.2 --CH═CH.sub.2                                         ( 10)

in which R¹, R², R³, X and Y have the above-given meanings, and, bysplitting off of the allyl ester group, this compound is converted intoa compound of the general formula (I), wherein A is a carboxyl group,and, if desired, in a compound of general formula (I), a radical A isconverted into another radical A within the above-given definition of A.

The acid XH is preferably an N-protected amino acid and the salt thereofis preferably a caesium salt.

The conditions for the individual, known reaction steps (temperature,solvent, period of reaction, etc.) correspond to the conditions usualfor such reactions. The reaction conditions which are most favourablefor individual cases can easily be determined and, as a rule, lie withinthe range of the reactions described in connection with FIGS. 1 to 3 ofthe accompanying drawings and within the reaction conditions given inthe following Examples.

FIGS. 1 to 3 of the accompanying drawings show the process according tothe present invention for the preparation of compounds of generalformula (I) using the example of representative compounds.

FIG. 1 shows the course of the reaction according to process variant a);

FIG. 2 shows the course of the reaction according to process variant b);and

FIG. 3 shows the course of the reaction according to process variant c).

According to process variant a) (FIG. 1), a salt of a compound HX isreacted with compound (2) in dimethylformamide, while stirring. Thecompound (3) obtained is dissolved in glacial acetic acid and convertedin the presence of the threefold amount of zinc into the compound (4).The coupling of (4) with (5) (in the form of the hydrochloride) takesplace by reacting a solution of (5) in methylene chloride/triethylamineor Hunig base with (4) in the presence N,N'-dicyclohexylcarbodiimide(DCC) and 1-hydroxybenzotriazole (HOBt) or also with the use of2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ). The conversion(ester cleavage) of the compound (I)(in which --R³ --A is the radical--(CH₂)_(n) --COO--CH₂ --CH₂ Br) into the compound (I)(in which --R³ --Ais the radical --(CH₂)_(n) --COOH) can take place in dimethylformamidein the presence of zinc/sodium iodide.

According to process variant b)(FIG. 2), the coupling of (6) with (5) togive (7) can take place analogously to the coupling of (4) with (5)described above under a), for example with DCC and EEDQ as couplingreagent. The reaction of (7) with the salt of the acid HX to give acompound of general formula (I) takes place analogously to the reactionof the salt of the acid HX with (2) described for process variant a).

According to process variant c)(FIG. 3), the coupling of (8) with (6) togive (9) takes place analogously to the corresponding reactions inprocess variant a) or b), for example with EEDQ as coupling reagent. Thereaction of (9) with the salt of the acid HX also takes placeanalogously to the corresponding reaction step in process variant a) orb). The cleavage of the ester (10) to give a compound of general formula(I) takes place, for example, in glacial aceticacid/tetrahydrofuran/water (20:20:1 v/v/v) with palladium chloride.

For the preparation of the compounds of general formula (I), as acid HXthere is preferably used an N-protected amino acid and, as salt, thecaesium salt is preferred.

As protective groups, there can be used the conventionally emplopyedprotective groups, for example Z (Cbo, Cbz), Boc, Ddz, Trt, Bpoc (Dpoc),OBut, Obzl, Nps, Fmoc, Msoc, Mch, Dcboc (with regard to theabbreviations, cf. for example Eur. J. Biochem., 74, 1-6/1977).

The present invention is also concerned with the use of a compound ofgeneral formula (I) according to the present invention for the build-upof a solid phase system of the general formula:

    F--(R.sup.3 --Y--CO--CR.sup.2 ═CR.sup.1 --CH.sub.2 --X).sub.m(II)

wherein F indicates a solid carrier material, R¹, R², R³, Y and X havethe above-given meanings and m is the number of side chains bound to thecarrier material; especially for use in solid phase reactions and, aboveall, for the solid phase syntheses of peptides, glycopeptides,nucleotides and proteins.

The solid phase systems of general formula (II) correspond to the solidphase systems described in published Federal Republic of Germany PatentSpecification No. 38 03 545. Because of their allylic side chain, thesesolid phase systems are suitable for the solid phase synthesis ofpeptides and glycopeptides in which the peptides and glycopeptides canbe split selectively from the polymeric carrier and under such mildconditions that no cleavage of the glycosidic bonds and noanomerisations take place.

The splitting off in the presence of catalytic amounts of a compound ofthe platinum group of metals, preferably in the presence of a rhodium(I) compound and especially in the presence of a palladium (O) compound,can be carried out in a solvent or solvent system appropriate for thispurpose, for example in tetrahydrofuran, in the presence of anucleophilic compound, for example morpholine, dimedone or anothereasily deprotonisable CH-acidic compound. The reaction is preferablycarried out at ambient temperature and is carried out with the exclusionof oxygen.

Because of the extraordinarily mild conditions of splitting off, withthe solid phase systems according to the present invention, it ispossible to carry out the splitting off of the peptides, glycopeptides,nucleotides and other radicals selectively and without cleavage ofglycosidic bonds or of anomerisations or isomerisations.

The present invention also provides a process for the preparation of asolid phase system of the above-given general formula (II), wherein acompound of general formula (I) according to the present invention isreacted with a solid carrier material F, which contains appropriatefunctional groups B, and wherein A and B signify atom groupings whichreact with one another with condensation and/or addition and formationof a linkage between the solid carrier material and the radical X--CH₂--CR¹ ═CR² --CO--Y--R³ --.

The solid carrier material is preferably an organic or inorganic polymeror also a solid base material which is coated with a material suitablefor linking with the compounds of general formula (I). As polymer or asmaterial suitable for linking with compounds of general formula (I),there is especially preferably used cross-linked polystyrene. Inparticular, the solid, functional group B-containing carrier materialsis an aminomethylated polystyrene.

The following Examples are given for the purpose of illustrating thepresent invention, the following abbreviations thereby being used:

As=amino acid

DCC=N,N'-dicyclohexylcarbodiimide

DIIC=N,N'-diisopropylcarbodiimide

EEDQ=2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline,

HOBt=1-hydroxybenzotriazole

SG=protective group.

EXAMPLE 1 Crotonic acid 2,2,2-trichloroethyl ester (i)

In 200 ml. carbon tetrachloride are dissolved 0.5 mole (43.07 g.)crotonic acid and the equimolar amount of 2,2,2-trichloroethanol, 1 ml.concentrated sulphuric acid is added thereto and the reaction mixture isheated under reflux on a water separator until the theoretical amount ofwater (9 ml.) has separated off. Subsequently, the organic phase isshaken out twice with, in each case, 100 ml. aqueous 2N sodium hydrogencarbonate solution and twice with, in each case, 50 ml. of water, theorganic phase is then dried with anhydrous magnesium sulphate and thesolvent is distilled off in a vacuum. There is finally obtained thedesired product (i) in the form of a bright yellow oil in a yield of 95%of theory. C₆ H₇ O₂ Cl₃ (217.4791)

Elementary analysis

calc.: C 33.13%; H 3.24%; Cl 48.90%; found: 32.97%; 3.43%; 48.82%; Rfvalue: 0.57 (PE/EE: 20:1 v/v).

200 MHz ¹ H-NMR (CDCl₃): 7.25-7.03, 1H, m, CH═, J_(trans) =15.6, J_(cis)=10.6, J=1.6 Hz; 5.98-5.88, 1H, m, CH═, J=7 Hz; 4.76, 2H, s, CH₂ ; 1.91,dd, 3H, CH₃.

50 MHz ¹³ C-NMR (CDCl₃): 164.55; C═O; 147.51; 121.25; CH═CH; 95.17; CCl₃; 73.84; CH₂ ; 18.26; CH₃.

EXAMPLE 2 4-Bromocrotonic acid 2,2,2-trichloroethyl ester (ii)

The bromination reaction takes place in carbon tetrachloride withN-bromosuccinimide and AIBN as radical former. For this purpose, anequimolar amount of ester (i) and 0.45 mole N-bromosuccinimide are mixedwith 300 ml. carbon tetrachloride and a spatula tip of AIBN addedthereto. The reaction mixture is heated under reflux for 4 hours,subsequently allowed to cool and the precipitated succinimide (0.41mole; 92% of theory) is filtered off. After distilling off the solventin a vacuum, the residue is purified by column chromatography usingPE/EE (25:1 v/v) as elution agent. The end product obtained is a yellowoil. The yield of 75% of theory. C₆ H₆ O₂ BrCl₃ (296.3752 g./mole).

Elementary analysis

calc.: C 24.31%; H 2.04%; Br 26.96%; Cl 35.88%; found: 24.61%; 1.99%;26.75%; 35.54%.

Rf value MHz ¹ H-NMR (CDCl₃): 7.25-7.06, 1H, m, CH═, J_(trans) =15.3 Hz,J_(cis) =7.3, J=6.3 Hz; 6.18-6.09, 1H, m, CH═; 4.79, 2H, s, CH₂ ; 4.03,dd, 3H, CH₂.

50 MHz ¹³ C-NMR (CDCl₃): 160.95; C═O; 141.48; 120.37; CH═CH; 92.17, CCl₃; 71.53; CH₂ ; 25.88; CH₂ Br.

EXAMPLE 3 N-tert.-butyloxycarbonylalanyloxycrotonic acid2,2,2-trichloroethyl ester (iii)

As precursor of this reaction, there first takes place the preparationof the caesium salt of the amino acid. For this purpose, 1.89 g. (0.01mole) of Bocalanine is dissolved in 10 ml. methanol and mixed with 5mmole caesium carbonate. With the evolution of carbon dioxide, there isformed the caesium salt which, after removal of the solvent bydistillation in a vacuum, is dissolved in dimethylformamide. 0.011 moleof (ii) is now added thereto and the reaction mixture is stirred for 12hours. Subsequently, precipitated caesium bromide is filtered off andthe dimethylformamide is distilled off in a vacuum. The crude product ispurified by column chromatography using, as elution agent, PE/EE (4:1v/v). Yellowish crystals are obtained. C₁₄ H₂₀ O₆ NCl₃ (404.6741g./mole)

Elementary analysis

calc.: C 41.55%; H 4.98%; N 3.46%; Cl 26.28%; found: 41.84%; 4.82%;3.31%; 26.38%; Rf value: 0.42 (PE/EE 4:1 v/v). Melting point: 58° C.[α]_(D) ²² =-21.26 (c=1, methanol)

200 MHz ¹ H-NMR (CDCl₃): 7.15-6.99, 1H, m, CH═, J_(trans) =15.5 Hz,J_(cis) =7.5, J=4.3 Hz; 6.16-6.08, 1H, m, CH═; 5.07, 1H, d, NH, J=6.3Hz; 4.77, 2H, s, CH₂ -ester; 4.88-4.72, 2H, m, CH₂ -crot., 4.37, 1H, dd,CH-Ala, J=7.1 Hz, 1.51-1.37, 12H, m, 4×CH₃.

50 MHz ¹³ C-NMR (CDCl₃): 172.66; 163.78; 155.03; C═O; 143.22; 120.71;CH═CH; 94.87; CCl₃ ; 80.08; C_(q) -Boc, 74.12; CH₂ ; 62.99; CH₂ -croton;49.35; CH-Ala; 28.30; CH₃ -Boc; 18.34; CH₃ -Ala.

EXAMPLE 4 N-tert.-Butyloxycarbonylalanyloxycrotonic acid (iv)

(cf. B. Marinier et al., Can. J. Chem., 51, 208-214/1973)

1.2 g. (2.96 mmole) of the ester from Example 3 is dissolved in 50 ml.glacial acetic acid and mixed with the threefold molar amount of zinc(9.17 mmole; 0.6 g.; previously activated with 1N hydrochloric acid).The reaction mixture is stirred for 2 hours and the acetic acidsubsequently removed by distilling off in a vacuum. The residue is nowtaken up in 50 ml. each of chloroform and saturated sodium hydrogencarbonate solution and filtered over a little kieselguhr. The organicphase is then extracted several times with 20 ml. amounts of sodiumhydrogen carbonate solution. The combined aqueous alkaline phases arecovered with ethyl acetate and carefully acidified with dilutehydrochloric acid to pH 2. Extraction is carried out three times with,in each case, 30 ml. ethyl acetate, the combined organic phases aredried with anhydrous magnesium sulphate and the solvent is removed bydistillation in a vacuum. If necessary, the product is further purifiedby means of column chromatography (elution agent PE/EE 1:1 v/v).Finally, there are obtained colourless crystals of (iv) in quantitativeyield. C₁₂ H₁₉ O₆ N (273.2852).

Elementary analysis

calc.: C 52.74%; H 7.00%; N 5.12%; found: 52.65%; 7.23%; 4.89%; Rfvalue: 0.21 (PE/EE 2:1 v/v); melting point: 65° C. [α]_(D) ²² =-36.12(c=1, methanol)

200 MHz ¹ H-NMR (CDCl₃); 7.25-6.93, 1H, m, CH═, J_(trans) =15.7 hz,J_(cis) =9.7, J=2.2 Hz; 6.06-5.98, 1H, m, CH═; 5.12, 1H, d, NH, J=7.3Hz; 4.78-6.06-5.98, 1H, m, CH═; 5.12, 1H, d, NH, J=7.3 Hz; 4.78-4.72,2H, m, CH₂ -crot., 4.37, 1H, dd, CH-Ala, J=7.3 Hz; 1.50-1.23, 12H, m,4×CH₃.

50 MHz ¹³ C-NMR (CDCl₃): 172.68; 169.89; 155.03; C═O; 142.66; 121.91;CH═CH; 80.08; C_(q) -Boc, 63.08; CH₂ ; 49.29; CH-Ala, 28.27; CH₃ -Boc;18.33; CH₃ -Ala.

EXAMPLE 5 Preparation of amino acid 2-bromoethyl ester hydrochlorides

(cf. H. Kunz, M. Buchholz, Chem. Ber., 112, 2145/1979)

0.2 mole of the amino acid is slurried, with stirring, in about 150 ml.freshly distilled 2-bromoethanol and cooled to -10° C. 30 g. (0.25 mole)distilled thionyl chloride are slowly added dropwise thereto, thereaction mixture is allowed to come to ambient temperature and thereaction mixture is stirred until a completely clear solution isobtained. The clear solution is slowly added dropwise, while stirring,into about 1 liter of diethyl ether/petroleum ether (2:1 v/v). Theprecipitated crystalline hydrochlorides are filtered off with suctionand subsequently washed with the precipitation agent. Excess halohydrinis recovered from the filtrate. All bromoethyl esters can be obtained ina yield greater than 98% of theory.

β-Alanine-2-bromoethyl ester hydrochloride

C₅ H₁₁ O₂ NBrCl (232.5044)

Elementary analysis

calc.: C 25.83%; H 4.77%; N 6.02%; Br 34.37%; Cl 15.25%; found: 25.76%;4.55%; 5.98%; 34.52%; 15.14%; Rf value: 0.21 (dichloromethane/methanol1:1 v/v); melting point: 68° C.

200 MHz ¹ H-NMR (DMSO-d₆): 7.99, 3H, s, NH₃ ⁺ ; 4.35, 2H, t, CH₂ ester,=5.5 Hz; 3.85, 2H, t, CH₂ -ester; 3.00, 2H, t, CH₂ -β-Ala, J=7 Hz; 2.74,2H, t, CH₂ -β-Ala.

50 MHz ¹³ C-NMR (DMSO-d₆): 169.83; C═O; 64.33; 42.28; 34.53; 31.21; CH₂.

6-Aminocapronic acid 2-bromoethyl ester hydrochloride

C₈ H₁₇ O₂ NBrCl (274.5852)

Elementary analysis

calc.: C 34.99%; H 6.24%; N 5.10%; Br 29.10%; Cl 12.91%; found: 34.56%;5.82%; 4.91%; 29.16%; 12.52%.

Rf value ¹ H-NMR (DMSO-d₆): 7.94, 3H, s, NH₃ ⁺ ; 4.25, 2H, t, CH₂-ester, J=5.5 Hz; 3.79, 2H, t, CH₂ -ester; 2.74, 2H, m, CH₂, J=6.1 Hz;2.27, m, CH₂, J=7.4 Hz; 1.62-1.14, 6H, m, CH₂.

50 MHz ¹³ C-NMR (DMSO-d₆): 172.41; C═O, 63.70; 42.53; 35.04; 30.79;26.39; 25.12; 23.75; CH₂.

EXAMPLE 6 N-tert.-Butyloxycarbonylalanyloxy-4-crotonyl-β-alanine2-bromoethyl ester (v)

9 mmole β-alanine 2-bromoethyl ester hydrochloride (va) (2.09 g.) aredissolved in 100 ml. anhydrous methylene chloride, the equivalent amountof triethylamine (0.91 g.) or of Hunig base are added thereto and thereaction mixture is stirred for 1 hour.

Into this solution are now added 2.46 of (iv) (9 mmole), 2.12 g. DCC (10mmole) and 2.03 g. HOBt (15 mmole). Already after a few minutes,dicyclohexylurea precipitates out of the reaction. However, forcompletion of the reaction, stirring is continued for 12 hours. Afterfiltering off the urea derivative and removing the solvent bydistillation in a vacuum, the crude product obtained is purified bychromatography, using ethyl acetate as elution agent.

The coupling can also be carried out with the help of EEDQ but theyields always lie between 44 and 59% of theory.

C₁₇ H₂₇ O₇ N₂ Br (451.3113)

Elementary analysis

calc.: C 45.24%; H 6.03%; N 6.21%; Br 17.70%; found: 45.26%; 6.41%;6.26%; 17.43%; Rf value: 0.62 (EE).

Melting point: 72°-73° C. [α]_(D) ²² =-19.84 (c=1, methanol)

200 MHz ¹ H-NMR (CDCl₃): 6.78-6.61, 2H, m, NH, CH═, J_(trans) =15.4 Hz,J_(cis) =9.2 Hz, J_(NH) =7.3 Hz; 6.09-5.94, 1H, m, CH═, J=4.3 Hz; 5.18,1H, d, NH, J=7.7 Hz; 4.69-4.67, 2H, m, CH₂ -crot.; 4.35-4.23, 3H, m,CH-Ala, J=7.3 Hz, CH₂ -ester, J=6 Hz; 3.52-3.40, 4H, m, CH₂ -ester, CH₂-β-Ala, J=6.1 Hz; 2.54, 2H, t, CH₂ -β-Ala; 1.34-1.19, 12H, m, 4×CH₃.

50 MHz ¹³ C-NMR (CDCl₃): 172.74; 171.64; 164.88; 155.15; C═O; 136.40;125.05; CH═CH; 79.97; C_(q) -Boc; 49.39; CH-Ala; 63.92; CH₂ -croton;63.44; 35.02, 33.92; 18.25; CH₂ ; 28.28; CH₃ -Boc; 17.87; CH₃ -Ala.

EXAMPLE 7 N-tert.-Butyloxycarbonylalanyloxy-4-crotonyl-β-alanine (vi)

The cleavage of the bromoethyl ester takes place analogously to H. Kunzand M. Buchholz (cf. H. Kunz, M. Buchholz, Chem. Ber., 112, 245/1979).

The bromoethyl ester (v)(3.5 mmole; 1.58 g.) is dissolved in 30 ml.dimethylformamide, a spatula tip of sodium iodide is added thereto andthe reaction mixture is stirred for 1 hour at 45° C. The threefold molaramount of activated zinc is now added thereto and the reaction mixtureis stirred at ambient temperature until the reaction is complete (about12 hours; TLC monitoring, PE/EE 2:1 v/v). After distilling off thesolvent in a vacuum, further working is carried out analogously to thatused for compound (iv). For the removal of catalyst residues (iodine),the organic phase is additionally shaken out three times with, in eachcase, 15 ml. 1N aqueous sodium thiosulphate solution. If, after theshaking out, the compound is still not pure (TLC monitoring), it ispurified by column chromatography (methanol/dichloromethane 3:1 v/v).There is finally obtained a bright yellow oil which, in the case of theaddition of diethyl ether/petroleum ether, precipitates out in the formof colourless crystals. The yield of this "triple-handled" compound is75% of theory.

C₁₅ H₂₄ O₇ N₂ (344.3652).

Elementary analysis

calc.: C 52.31%; H 7.02%; N 8.13%; found: 52.25%; 7.07%; 8.26%; Rfvalue: 0.22 (EE); 0.66 (methanol/dichloromethane 3:1 v/v).

Melting point: 43°-45° C. [α]_(D) ²² =-15.54 (c=0.99, methanol)

400 MHz ¹ H-NMR (CDCl₃): 10.1, 1H, s, COOH; 6.97, 1H, d, NH-Ala, J=7.3Hz; 6.97-6.72, 1H, m, CH═, J_(trans) =15.4 Hz; 6.05-6.04, 1H, m, CH═,J=4.6 Hz; 5.31, 1H, d, NH, J=7.3 Hz; 4.70-4.69, 2H, m, CH₂ -crot; 4.26,1H, dd, CH-Ala, J=7.3 Hz; 3.51, 2H, t, CH₂ -β-Ala, J=6.1 Hz; 2.53, 2H,t, CH₂ -β-Ala; 1.39-1.32, 12H, m, 4×CH₃.

100 MHz ¹³ C-NMR (DMSO-d₆): 175.35; 172.54; 163.80; 155.17; C═O; 134.91;125.84; CH═CH; 78.10, C_(q) -Boc; 49.91; CH-Ala; 62.97; CH₂ -croton.;36.25; 35.90; CH₂ ; 28.08; CH₃ -Boc; 16.83; CH₃ -Ala.

EXAMPLE 8 N-tert.-Butyloxycarbonylalanyloxy-4-crotonyl-6-amidocaproicacid 2-bromoethyl ester (vii)

The preparation of this compound does not take place as described inExample 6 with DCC as coupling reagent but with the help of EEDQ.Otherwise, the batch and course of the reaction are analogous. Aftercolumn chromatography with PE/EE (2:1 v/v), there is obtained ayellowish oil which, upon the addition of diethyl ether/petroleum ether,crystallises in colourless form. The yield is 69% of theory. C₂₀ H₃₃ O₇N₂ Br (493.3953)

Elementary analysis

calc.: C 48.68%; H 6.74%; N 5.67%; Br 16.19%; found: 48.89%; 6.91%;5.41%; 15.88%; Melting point: 58° C. [α]_(D) ²² =-11.99 (c=1, methanol)

400 MHz ¹ H-NMR (DMSO-d₆): 8.0, 1H, t, NH-capr., J=5.6 Hz; 7.3, 1H, d,NH-Ala, J=7.3 Hz; 6.60-6.53, 1H, m, CH═, J_(trans) =15.5 Hz, J_(cis)=9.7 Hz, J=1.6 Hz, 6.05-6.03, m, 1H, M, CH═, J=4.8 Hz, 4.75-4.67, 2H, m,CH₂ -crot.; 4.32, 2H, t, CH₂ -ester, J=5.7 Hz; 4.05, 1H, dd, CH-Ala,J=7.3 Hz; 3.65, 2H, t, CH₂ -ester; 3.08, 2H, m, CH₂, J=6.1 Hz; J=6.8 Hz;2.33-2.29, 2H, m, CH₂, J=7.4 Hz; 1.56-1.25, 18H, m, 4×CH₃, 3×CH₂.

100 MHz ¹³ C-NMR (DMSO-d₆): 172.57; 172.39; 163.87; 155.18; C═O, 135.12;125.52; C═C, 78.12; C_(q) -Boc; 63.43; 30.63; CH₂ -bromoethyl ester;62.95; CH₂ -croton.; 48.92; CH-Ala; 38.87; 33.22; 28.62; 25.75; 24.04;CH₂ ; 28.08; CH₃ -Boc; 16.84; CH₃ -Ala.

EXAMPLE 9 N-tert.-Butyloxycarbonylalanyloxy-4-crotonyl-6-amidocaproicacid (viii)

The "C-terminal handle" is obtained analogously to the process describedin Example 7 by cleavage of the corresponding bromoethyl ester (vii).Already after 9 hours, the thin layer chromatogram shows a completereaction to a uniform product. After working up by acidic and alkalineshaking up analogously to Examples 7 and 4, respectively, there isobtained, in a yield of 53% of theory, the C-terminal free compound inthe form of a yellowish oil which, already like the bromoethyl ester,precipitates out in the form of colourless crystals upon the addition ofdiethyl ether/petroleum ether.

C₁₈ H₃₀ O₇ N₂ (386.4456)

Elementary analysis

calc.: C 55.94%; H 7.82%; N 7.24%; found: 55.83%; 7.87%; 7.35%; Rfvalue: 0.20 (EE); Melting point: 71°-76° C. [α]_(D) ²² =-17.64 (c=1,methanol)

400 MHz ¹ H-NMR (DMSO-d₆): 11.99, 1H, s, COOH; 8.05, 1H, t, NH-capr.,J=5.4 Hz; 7.31, 1H, d, NH-Ala, J=7.3 Hz; 6.60-6.53, 1H, m, CH═,J_(trans) =15.5 Hz, J_(cis) =9.7 Hz, J=1.6 Hz; 6.05-6.03, m, 1H, m, CH═,J=4.8 Hz; 4.75-4.64, 2H, m, CH₂ -crot.; 4.05, 1H, dd, CH-Ala, J=7.3 Hz;3.17-3.05, 2H, m, CH₂, J=6.1 Hz, J=6.7 Hz; 2.19-2.15, 2H, m, CH₂, J=7.4Hz; 1.51-1.19, 18H, m, 4×CH₃, 3×CH₂.

100 MHz ¹³ C-NMR (DMSO-d₆): 174.33; 172.64; 163.95; 155.25, C═O, 135.21,125.59; C═C; 78.17; C_(q) -Boc; 62.98; CH₂ -crotonic acid; 48.95;CH-Ala; 38.36; 33.53; 28.71; 25.93; 24.14; CH₂ ; 28.10; CH₃ -Boc; 16.85;CH₃ -Ala.

EXAMPLE 10 Synthesis of the N-protected pentapeptideZ-Val-Ala-Leu-Gly-Ala-OH

This is prepared with the use of the "handles" from Examples 7 and 9 andsplitting off of the peptide from the carrier resin.

The synthesis of the pentapeptide takes place under standardisedconditions with a peptide synthesiser PSS 80 of the firm Applied ProteinTechnics.

The individual reaction steps are as follows: In each case, 489 mg.p-aminomethylpolystyrene with the loading of 1.7 mmole NH₂ /g. of resin(thus here corresponds to 0.84 mmole anchor groups) are suspended in 10ml. of anhydrous dimethylformamide and mixed with the 4 fold molaramount of the corresponding "triple handle" (vi) and (viii),respectively (3.4 mmole), 4 mmole DIC and 4 mmole HOBt. After 25minutes, the resin is washed with dimethylformamide and the couplingstep is repeated. An internal monitoring examines the reaction. Afterthe second coupling, a capping step takes place with acetic anhydride(20 minutes, 10 ml. 30% acetic anhydride in dimethylformamide). Thereaction is subsequently examined by qualitative and quantitative aminoacid sequence analysis. Subsequently, there takes place the splittingoff of the Boc protective group with 20 ml. TFA/dichloromethane (1:1v/v; 1 hour) and, after neutralisation and washing of the resin withtriethylamine or dimethylformamide, the elongation of the peptide to thedesired length. The coupling reagent is again DIC/HOBt and the Fmocgroup is used as N-terminal protective group, in the case of valine theZ group. The molar excess is always four and coupling is carried outtwice. The splitting off of the Fmoc protective group takes placethroughout with piperidine and, in each case, the coupling period is 25minutes. For the splitting off of the protective group, in each case 1hour is necessary. After completion of the synthesis to give theN-terminal (Z)-protected pentapeptide, a quantitative and qualitativeamino acid sequence analysis is carried out in order to monitor thesynthesis and to obtain the loading of the resin with peptide. Theβ-alanine used as standard is thereby detected as phenylalanine(retention time: 46.883 minutes); capric acid is superimposed withhistidine (the retention time is here 49.237 minutes).

The peptide is now split off from the polymeric carrier. For thispurpose, the peptide-carrying resin (depending upon the reaction about700 mg.) is introduced into anhydrous and oxygen-freetetrahydrofuran/dichloromethane/dimethyl sulphoxide (1:1:1 v/v/v), usingargon as protective gas, three times the theoretical amount ofmorpholine are added thereto (referred to a quantitative reaction: 2.6mmole) and mixed with a spatula tip of catalyst(tetrakis(triphenylphosphine)-palladium-(O) of the firm Janssen).Shaking is then carried out with the exclusion of light and oxygen for12 hours, using argon as protective gas.

After completion of the cleavage reaction, the carrier resin isseparated off and again washed three times with, in each case, 30 ml.tetrahydrofuran and methylene chloride. The solvent is removed bydistillation from the combined organic filtrates and the residue ispurified by column chromatography. The elution agent used is ethylacetate/isopropanol/glacial acetic acid (12:1:0.1 v/v/v). There arefinally obtained pale yellowish crystals.

Analytical data of the pentapeptide: C₂₇ H₄₁ O₈ N₅ (563.648)

Amino acid sequence analysis (hydrochloric acid/propionic acid; 110° C.,caproic acid as internal standard):

calc.: Gly=1; Ala=2; Leu=1; Val=1; found: Gly=1.2; Ala=1.85; Leu=1.09;Val=0.84.

Amino acid sequence analysis (hydrochloric acid/propionic acid; 110° C.,β-alanine as internal standard):

calc.: Gly=1; Ala=2; Leu=1; Val=1; found: Gly=1.1; Ala=2.09; Leu=0.93;Val=0.9 Rf value: 0.62 (EE/isopropanol/HAc=12:1:0.1 v/v/v)

Melting point: 141°-144° C. [α]_(D) ²² =0.31 (c=1, methanol) 100 MHz ¹³C-NMR (DMSO-d₆)

Analysis data for the synthesis of the pentapeptide:

The loading of the resin was, in each case, determined by means ofquantitative amino acid sequence analysis (conditions see above) andreferred to the particular internal standard.

a) Internal standard β-alanine:

Loading of the resin with "triple handle": 0.6 mmole/g.; 36% amino acidsequence analysis after the synthesis to the dipeptide (resin-bound;Fmoc-Gly-Ala):

calc.: Gly=1; Ala=1; found: Gly=0.94; Ala=1.05. Loading of the resinwith pentapeptide: 0.6 mmole/g.; 36%. Theoretically cleavable amount ofpeptide (489 mg. of resin were used): 169 mg. (0.3 mmole) Yield ofpeptide cleaved off: 142 mg. (0.25 mmole) Yield of peptide referred tothe loading of the resin with "triple handle": 83%. Yield of peptidereferred to the theoretically possible amount: 30%.

b) Internal standard 6-aminocaproic acid:

Loading of the resin with "triple handle": 1.1 mmole/g.; 65% Amino acidsequence analysis after the synthesis to the dipeptide (resin-bound;Fmoc-Gly-Ala):

calc.: Gly=1; Ala=1; found: Gly=0.97; Ala=1.2

Amino acid sequence analysis after the synthesis to the tetrapeptide(resin-bound; Fmoc-Ala-Leu-Gly-Ala):

calc.: Gly=1; Ala=2; Leu=1; found: Gly=1.09; Ala=1.77; Leu=1.13; Loadingof the resin with pentapeptide: 1.1 mmole/g., 65%; Theoreticallycleavable amount of peptide (489 g. of resin were used): 303 mg. (0.54mmole); Yield of peptide cleaved off: 293 mg. (0.52 mmole) Yield ofpeptide referred to the loading of the resin with "triple handle": 97%.Yield of peptide referred to the theoretically possible amount: 62%.

We claim:
 1. Process for the preparation of a solid phase system of theformula:

    F--(--R.sup.3 --Y--CO--CR.sup.2 ═CR.sup.1 --CH.sub.2 --X).sub.m(II)

wherein R¹ and R² which can be the same or different are hydrogen,halogen, C₁ -C₇ alkyl or mono or dicyclic aryl, R³ is a linking group(CH₂)_(n) COOCH₂ CH₂ or (CH₂)_(n) wherein n is 2-10, X is acyloxywherein the acyl is the residue of a C₁ -C₇ aliphatic carboxylic acid oran N-protected amino acid or is RCO-- wherein R is the protected orunprotected residue of an amino acid, peptide, glycopeptide, nucleotide,hydroxy carboxylic acid, dicarboxylic acid or tricarboxylic acid and Yis oxygen, sulphur, or an --NH--, A is a carboxylic acid ester groupcapable of reaction with a functional group B present on a solid carriermaterial selected from the group consisting of cross-linkedpolyacrylamides, methacrylates, dextrans, cellulose, polystyrene andcross-linked polystyrene, F is a solid phase carrier material and m isthe number of side chains bound to the carrier material comprisingreacting an allyl ester of formula I:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --A  (I)

with a solid phase carrier-material F containing functional groups Bwherein R¹, R², R³, X and Y have the above meanings and wherein A and Breact with one another to form a linkage between the solid phase carriermaterial F and X--CH₂ --CR¹ ═CR² --COY--R³ comprising reacting acompound of formula

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --COOOH.sub.2 CCl.sub.3( 2)

in which R¹ and R² have the above-given meanings, is reacted with a saltof an acid of the general formula XH (1), in which X has the above-givenmeaning, the so-obtained trichloroethyl ester of formula:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 COOH.sub.2 Cl.sub.3   ( 3)

in which R¹, R² and X have the above-given meanings, is converted intothe free acid of formula:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --COOH                (4)

in which R¹, R² and X have the above-given meanings, and this is reactedwith a compound of formula:

    HY--R.sup.3 --A                                            (5)

in which A, R³ and Y have the above-given meanings, to give a compoundof formula (I).
 2. Process for the preparation of a solid phase systemof formula:

    F--(--R.sup.3 --Y--CO--CR.sup.2 ═CR.sup.1 --CH.sub.2 --X).sub.m(II)

wherein R¹ and R² which can be the same or different are hydrogen,halogen, C₁ -C₇ alkyl or mono or dicyclic aryl, R³ is a linking group(CH₂)_(n) COOCH₂ CH₂ or (CH₂)_(n) wherein n is 2-10, X is acyloxywherein the acyl is the residue of a C₁ -C₇ aliphatic carboxylic acid oran N-protected amino acid or is RCO-- wherein R is the protected orunprotected residue of an amino acid, peptide, glycopeptide, nucleotide,hydroxy carboxylic acid, dicarboxylic acid or tricarboxylic acid and Yis oxygen, sulphur, or an --NH--, A is a carboxylic acid ester groupcapable of reaction with a functional group B present on a solid carriermaterial selected from the group consisting of cross-linkedpolyacrylamides, methacrylates, dextrans, cellulose, polystyrene andcross-linked polystyrene, F is a solid phase carrier material and m isthe number of side chains bound to the carrier material comprisingreacting an allyl ester of formula I:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --A  (I)

with a solid phase carrier-material F containing functional groups Bwherein R¹, R², R³, X and Y have the above meanings and wherein A and Breact with one another to form a linkage between the solid phase carriermaterial F and X--CH₂ --CR¹ ═CR² --COY--R³ comprising reacting acompound of the formula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --COOH                 (6)

in which R¹ and R² have the above-given meanings with a compound offormula HY--R³ --A (5) to give a compound of the formula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --A   (7)

in which A, R¹, R² and Y have the above-given meanings, and convertingcompound (7) into a compound of formula (I) by reaction with a salt ofan acid of general formula XH (1), in which X has the above meaning. 3.Process for the preparation of a solid phase system of the generalformula:

    F--(--R.sup.3 --Y--CO--CR.sup.2 ═CR.sup.1 --CH.sub.2 --X).sub.m(II)

wherein R¹ and R² which can be the same or different are hydrogen,halogen, C₁ -C₇ alkyl or mono or dicyclic aryl, R³ is a linking group(CH₂)_(n) COOCH₂ CH₂ or (CH₂)_(n) wherein n is 2-10. X is acyloxywherein the acyl is the residue of a C₁ -C₇ aliphatic carboxylic acid oran N-protected amino acid or is RCO-- wherein R is the protected orunprotected residue of an amino acid, peptide, glycopeptide, nucleotide,hydroxy carboxylic acid, dicarboxylic acid or tricarboxylic acid and Yis oxygen, sulphur, or an --NH--, A is a carboxylic acid ester groupcapable of reaction with a functional group B present on a solid carriermaterial selected from the group consisting of cross-linkedpolyacrylamides, methacrylates, dextrans, cellulose, polystyrene andcross-linked polystyrene, F is a solid phase carrier material and m isthe number of side chains bound to the carrier material comprisingreacting an allyl ester of formula I:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --A  (I)

with a solid phase carrier-material F containing functional groups Bwherein R¹, R², R³, X and Y have the above meanings and wherein A and Breact with one another to form a linkage between the solid phase carriermaterial F and X--CH₂ --CR¹ ═CR² --COY--R³ comprising reacting acompound of the formula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --COOH                 (6)

with a compound of the formula:

    HY--R.sup.3 --COO--CH.sub.2 --CH═CH.sub.2              ( 8)

in which R³ and Y have the above-given meanings, to give a compound ofthe formula:

    BrCH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --COO--CH.sub.2 --CH═CH.sub.2                                         ( 9)

in which R¹, R², R³ and Y have the above-given meanings, reacting thecompound (9) with the salt of an acid of the formula XH (1), in which Xhas the above-given meaning, to give a compound of the formula:

    X--CH.sub.2 --CR.sup.1 ═CR.sup.2 --CO--Y--R.sup.3 --COO--CH.sub.2 --CH═CH.sub.2                                         ( 10)

in which R¹, R², R³, X and Y have the above-given meanings, andsplitting off the allyl ester group thereby converting the compound (10)into a compound of the formula (I), wherein A is carboxyl group.
 4. Themethod of claim 1, 2 or 3 wherein XH is an N-protected amino acidwherein the salt is a caesium salt.
 5. Process according to any one ofclaims 1, 2 or 3, wherein the solid carrier material is a solid basematerial which is coated with a material appropriate for linking withallyl esters of general formula (I).
 6. Process according to claim 5,wherein the solid phase, functional group B-containing carrier materialF is an aminomethylated polystyrene or polyacrylamide which is reactedwith an allyl ester of general formula (I) with the formation of anamide group.